Precise DNA cleavage using CRISPR-SpRYgests

The ability to precisely cleave DNA at specifiable bases is critical for many applications in life science research, including for molecular biology and cloning, genome editing, DNA sequencing, protein engineering, and a wide variety of other methods. Until now, no enzyme or method existed that permitted efficient cleavage of any position of a DNA substrate. Current workflows typically rely on restriction enzymes (REs) that are beholden to constraining 6-8 bp binding motifs, and yet despite a diverse catalog of REs, collectively they can only target a small fraction of all DNA sequences. To solve this key challenge, CGM Investigator Ben Kleinstiver and colleagues optimized our recently engineered RNA-programmed PAMless Cas9 enzyme, named SpRY, as a highly precise DNA cleavage tool. The use of SpRY for DNA digests (SpRYgests) enables precise manipulation of nucleic acids in ways not possible with REs or other nucleases. The applications of SpRYgests are vast and hold promise to expedite and improve a range of biomedical research endeavors.

Read more in a Tweetorial here, and read the preprint on BioRxiv here.

June 21, 2023


CGM Primary Investigator